Solid-state electrochemiluminescence protein biosensor with aptamer substitution strategy

One solid-state electrochemiluminescence （ECL） protein biosensor based on the competing reaction and substitute reaction between protein-to-DNA aptamer and DNA-to-DNA aptamer was proposed. Additionally, the biosensor was based on ECL photo-quenching effect of ferrocene （Fc） to tris（2,2-bipyridyl）ruthenium（II） （Ru（bpy）2＋）. It was built up by modification of Au nanoparticles （AuNPs） and Ru（bpy）32＋ on one Au electrode firstly, and then self-assembly of one special double-stranded DNA （dsDNA） onto the electrode. This dsDNA was prepared by hybridization of one Fc labeled molecular beacon single-stranded DNA（ssDNA） and one anti-thrombin aptamer ssDNA. Without the target protein, this Fc-dsDNA/Ru（bpy）2＋- AuNPs/Au elec- trode trigged strong ECL signal, so we called it ECL ＂signal on＂ state. When thrombin was present in the sensing solution, the protein reacted with its aptamer from the Fc-dsDNA/Ru（bpy）3^2＋-AuNPs/Au electrode. Then the left molecular beacon ssDNA on the electrode recovered to its normal stem-loop structure and consequently its Fc labeler was close enough to the electrode surface to quench the ECL signal from Ru（bpy）3^2＋. It was in ECL ＂signal off＂ state. We measured the decrease in ECL intensity to sense the target protein. This was one endeavour to sense protein by using un-labeling target or probe strategy, which gave higher sensitivity and selectivity due to the better combination efficiency of protein and the un-labeled aptamer. 6.25 fmo/L thrombin was detected out,

Recent Advances in Visual Electrochemiluminescence Analysis

Due to the advantages of low background interference and high sensitivity, electrochemiluminescence (ECL)-based sensor has developed rapidly in recent years. The ECL sensors have shown the potential in the ultrasensitive and real-time analysis. Especially, the visual ECL analysis, including visual detection, cell imaging and single particle analysis, has offered many unique sensing platforms for analysis research and point-of-care testing. The high throughput ECL image analysis can not only provide ECL intensity but also reveal more information about the chemical reaction activity of particle and the physi-ological processes of cell operation. Therefore, we review the novel ECL luminophore, sensing systems, and successful applications in the visual ECL detection and imaging in this paper. First, the different ECL luminophore is summarized. Second, we discuss the ECL sensing mechanisms, focusing on the advantages and limitations of different sensing methods. Then, we highlight the recent advances in representative examples of visual ECL analysis, including aptasensing, multiplex immunoassays, cell imaging and single-particle analysis. At last, the outlook and prospects for the future visual ECL analysis are discussed based on the current development of ECL research.

A nano-functionalized real-time electrochemiluminescent biosensor for alanine transaminase assay

An electrochemiluminescent (ECL) biosensor was constructed for selective assay of alanine aminotransferase (ALT) based on the enzymatically catalyzed oxidation of pyruvate by pyruvate oxidase (PYOD). The composite of potassium ferricyanide and carbon nanotube was adopted to pre-functionalize the basal platinum electrode while the potassium ferricyanide acted as the activator of PYOD. The ALT catalyzed the reaction of L-alanine and-ketoglutarate to produce pyruvate which could be further enzymatically oxidized by PYOD to yield H2O2 to intensify the ECL of luminol. The biosensor showed rapid response for real-time measurement of ALT in the linear concentration range from 0.00475 to 350 U/L (r = 0.993) with a relatively standard deviation of 2.5% (CALT = 47.5 U/L,n = 6). The biosensor was applied to assay the ALT in rat serum with average recovery of 90.5%.